Detection of organisms using a media sachet and primer directed nucleic acid amplification

Abstract

A method of detecting a target organism in a sample mixture comprising preparing at least one complex sample mixture, performing primer directed nucleic acid amplification of the complex sample mixture; and examining the primer directed nucleic acid amplification result to detect for presence or absence of amplification product of the target organism, wherein the presence or absence of the amplification product is indicative of the presence or absence of the target organism in the sample mixture. Preparing the complex sample mixture comprises adding to a container at least one sachet containing at least a nutrient concentrate and a sample mixture suspected of containing the target organism.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of U.S. Provisional Application No. 60 / 668,020, filed Apr. 4, 2005 and U.S. Provisional Application No. 60 / 708,037, filed Aug. 12, 2005, each of which is hereby incorporated by reference in its entirety.FIELD OF THE INVENTION [0002] The field of the invention relates to molecular biology and particularly to methods and compositions for preparing samples and detecting target organisms contained therein. BACKGROUND [0003] Nucleic acid based methods, and in particular polymerase chain reaction (PCR) methods, are powerful analytical tools in the detection and identification of organisms such as bacteria. Nucleic acid based methods include the amplification of any desired specific nucleic acid sequence contained in a nucleic acid or mixture thereof. The use of these methods for bacterial detection in known in the art. However, these methods typically demand adherence to strict protocols under strict condit...

AI Technical Summary

Benefits of technology

The present invention is a method for detecting a target organism in a sample mixture. The method involves preparing a complex sample mixture by adding a nutrient concentrate and a sample mixture containing the target organism to a container. The sample mixture is then subjected to nucleic acid amplification using primers directed towards the target organism. The amplification result is examined to detect the presence or absence of the target organism in the sample mixture. This method provides a reliable and sensitive way to detect target organisms in complex samples.

Problems solved by technology

However, these methods typically demand adherence to strict protocols under strict conditions and often require personnel of advanced skills and training in order to achieve a reliable result.

In all nucleic acid based methods for detection of organisms, and particularly in a PCR-based test procedure, extraneous components that may enhance or inhibit the test reaction make it difficult to obtain credible results.

However, media preparation at each testing laboratory is often expensive, labor intensive, and subject to error, especially during the measuring and transferring operations.

The food and environmental testing industry in particular considers this process of media preparation, sterilization, storage, and introduction, to be burdensome.

The availability of liquid media solutions, even in flexible medium bags, is still often undesirable, due to shipment drawbacks arising from the weight and volume of the solutions.